Characterization of PINK1 processing, stability, and subcellular localization.
Characterization of PINK1 processing, stability, and subcellular localization.
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DOI：
10.1111/j.1471-4159.2008.05398.x
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发表时间：
2008-07
影响因子：
4.7
通讯作者：
Kang UJ
中科院分区：
文献类型：
--
作者：
Lin W;Kang UJ
在PTEN诱导的推定激酶1（PINK 1）（一种功能未知的推定线粒体丝氨酸/苏氨酸激酶）中发现的突变与常染色体隐性帕金森病有关。这表明突变可导致PINK 1激酶活性丧失，并最终导致线粒体功能障碍。在这份报告中，我们研究了PINK 1的亚细胞定位和PINK 1加工和降解的动态动力学。我们还确定了胞质伴侣热休克蛋白90（Hsp 90）作为一个相互作用的蛋白质PINK 1的PINK 1免疫共沉淀。PINK 1蛋白的免疫荧光和线粒体分离显示PINK 1的前体形式易位到线粒体，并被加工成两种切割形式的PINK 1，这反过来又更多地定位于细胞溶质而不是线粒体部分。当线粒体膜电位被破坏时，不发生切割并且未切割的前体保持与线粒体缔合。代谢标记分析表明，PINK 1加工迅速，切割形式的水平受到严格调控。此外，裂解形式的PINK 1通过Hsp 90相互作用稳定，因为Hsp 90活性的丧失降低了线粒体加工后的PINK 1水平。最后，我们还发现PINK 1的切割形式被蛋白酶体降解，这对于线粒体蛋白是不常见的。我们的研究结果支持双重亚细胞定位，这意味着PINK 1可以驻留在线粒体和细胞质中。这就提出了连接这两个细胞区室的有趣的功能作用。
Mutations found in PTEN-induced putative kinase 1 (PINK1), a putative mitochondrial serine/threonine kinase of unknown function, have been linked to autosomal recessive Parkinson’s disease. It is suggested that mutations can cause a loss of PINK1 kinase activity and eventually lead to mitochondrial dysfunction. In this report, we examined the subcellular localization of PINK1 and the dynamic kinetics of PINK1 processing and degradation. We also identified cytosolic chaperone heat-shock protein 90 (Hsp90) as an interacting protein of PINK1 by PINK1 co-immunoprecipitation. Immunofluorescence of PINK1 protein and mitochondrial isolation show that the precursor form of PINK1 translocates to the mitochondria and is processed into two cleaved forms of PINK1, which in turn localize more to the cytosolic than mitochondrial fraction. The cleavage does not occur and the uncleaved precursor stays associated with the mitochondria when the mitochondrial membrane potential is disrupted. Metabolic labeling analyses show that the PINK1 processing is rapid and the levels of cleaved forms are tightly regulated. Furthermore, cleaved forms of PINK1 are stabilized by Hsp90 interaction as the loss of Hsp90 activity decreases PINK1 level after mitochondrial processing. Lastly, we also find that cleaved forms of PINK1 are degraded by the proteasome, which is uncommon for mitochondrial proteins. Our findings support a dual subcellular localization, implying that PINK1 can reside in the mitochondria and the cytosol. This raises intriguing functional roles that bridge these two cellular compartments.
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